LOCUS       sequence-231454-        8284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual luciferase assay for miRNA-targeted UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8284)
  AUTHORS   Kutsche LK, Gysi DM, Fallmann J, Lenk K, Petri R, Swiersy A, Klapper
            SD, Pircs K, Khattak S, Stadler PF, Jakobsson J, Nowick K, Busskamp 
            V
  TITLE     Combined Experimental and System-Level Analyses Reveal the Complex 
            Regulatory Network of miR-124 during Human Neurogenesis.
  JOURNAL   Cell Syst. 2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 
            10.1016/j.cels.2018.08.011.
  PUBMED    30292704
REFERENCE   2  (bases 1 to 8284)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Syst. 
            2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 
            10.1016/j.cels.2018.08.011."
COMMENT     Sequence Label: pmiRGLO-SEMA5B-211
FEATURES             Location/Qualifiers
     source          1..8284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     300..319
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      408..417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             414..2066
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    3168..3289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3179..3198)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3233..3252
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3534..3891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3534..3725
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3557..3577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3742..3877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3936..3945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3942..4874
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4926..5717
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4977..4996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5587..5606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5781..5829
                     /note="synthetic polyadenylation signal"
     promoter        5985..6089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6090..6158,6159..6950)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6308..6327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7108..7696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7188..7207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7812..8095
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(8152..8284,1..367)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8187..8210)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
       61 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
      121 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
      181 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
      241 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
      301 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
      361 cctttcgacc tgcagcccaa gcttggcaat ccggtactgt tggtaaagcc accatggaag
      421 atgccaaaaa cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg
      481 gcgagcagct gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta
      541 ccgacgcaca tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc
      601 tggcagaagc tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg
      661 agaatagctt gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg
      721 ccccagctaa cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc
      781 ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc
      841 taccgatcat acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa
      901 gcatgtacac cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc
      961 ccgagagctt cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg
     1021 gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc
     1081 gcgaccccat cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat
     1141 ttcaccacgg cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg
     1201 tgctcatgta ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc
     1261 aatctgccct gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca
     1321 agtacgacct aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg
     1381 taggtgaggc cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga
     1441 cagaaacaac cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag
     1501 gcaaggtggt gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg
     1561 gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta
     1621 acaaccccga ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca
     1681 tcgcctactg ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca
     1741 aatacaaggg ctaccaggta gccccagccg aactggagag catcctgctg caacacccca
     1801 acatcttcga cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg
     1861 cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg
     1921 ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc
     1981 ctaaaggact gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca
     2041 agaagggcgg caagatcgcc gtgtaattct agttgtttaa acgagctcgt accgccgtcc
     2101 tggggacttg ggcttcttgc cttcataagg cacagagcag atggagatgg gacagtggag
     2161 ccagtttggt tttctccctc tgcactaggc caagaacttg ctgccttgcc tgtggggggt
     2221 cccatccggc ttcagagagc tctggctggc attgaccatg ggggaaaggg ctggtttcag
     2281 gctgacatat ggccgcaggt ccagttcagc ccaggtctct catggttatc ttccaaccca
     2341 ctgtcacgct gacactatgc tgccatgcct gggctgtgga cctactgggc atttgaggaa
     2401 ttggagaatg gagatggcaa gagggcaggc ttttaagttt gggttggaga caacttcctg
     2461 tggcccccac aagctgagtc tggccttctc cagctggccc caaaaaaggc ctttgctaca
     2521 tcctgattat ctctgaaagt aatcaatcaa gtggctccag tagctctgga ttttctgcca
     2581 gggctgggcc attgtggtgc tgccccagta tgacatggga ccaaggccag cgcaggttat
     2641 ccacctctgc ctggaagtct atactctacc cagggcatcc ctctggtcag aggcagtgag
     2701 tactgggaac tggaggctga cctgtgctta gaagtccttt aatctgggct ggtacaggcc
     2761 tcagccttgc cctcaatgca cgaaaggtgg cccaggagag aggatcaatg ccataggagg
     2821 cagaagtctg gcctctgtgc ctctatggag actatcttcc agttgctgct caacagagtt
     2881 gttggctgag acctgcttgg gagtctctgc tggcccttca tctgttcagg aacacacaca
     2941 cacacacact cacacacgca cacacaatca caatttgcta cagcaacaaa aaagacattg
     3001 ggctgtggca ttattaatta aagatgatat ccagtctcca gtcgacctgc aggcatgcaa
     3061 gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     3121 caataactag cataacccct tggggcggcc gcttcgagca gacatgataa gatacattga
     3181 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3241 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3301 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3361 aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct
     3421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     3481 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3541 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3721 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3781 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3841 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     3901 tcttctgaca caacagtctc gaaccaaagg ctggagccac catggcttcc aaggtgtacg
     3961 accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa
     4021 tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc gagaacgccg
     4081 tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca
     4141 tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag tccggcaaga
     4201 gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct tggttcgagc
     4261 tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct tgtctggcct
     4321 ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct gagagtgtcg
     4381 tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc gccctgatca
     4441 agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag accatgctcc
     4501 caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg gagccattca
     4561 aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc cctctcgtta
     4621 agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac cttcgggcca
     4681 gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc aacgctattg
     4741 tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc ctccacttca
     4801 gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc
     4861 tgaagaacga gcagaccggt ggtgggagcg gaggtggcgg atcaggtggc ggaggctccg
     4921 gagggattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5101 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5461 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5701 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5761 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5821 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     5881 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     5941 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     6301 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6481 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     6541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     6961 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7681 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7801 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     7861 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     7921 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     7981 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     8041 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     8101 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatctac cgggtagggg
     8161 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
     8221 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
     8281 ctcc
//
